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Specific PCR primers for identifying rhodobacter capsulatus, kit and application

A capsulated rhodobacter, specific technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of long detection time and poor specificity, and achieve high sensitivity and good false detection Positive, solving the effect of a single detection target

Active Publication Date: 2019-09-17
HUNAN AGRICULTURAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Based on this, aiming at the technical problems of the above-mentioned method for identifying Rhodobacter capsularis in the above-mentioned prior art, which has complicated operation steps, long detection time and poor specificity, the present invention provides a primer, which has strong specificity for Rhodobacter capsularis, and can be used in During the identification process, the operation is simple, the detection time is short, the sensitivity is high, and the stability is strong

Method used

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  • Specific PCR primers for identifying rhodobacter capsulatus, kit and application
  • Specific PCR primers for identifying rhodobacter capsulatus, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0026] see figure 1 , identify the method for capsular rhodobacter, comprising the steps:

[0027] 1. Extraction of colony DNA to be tested

[0028] Weigh 1.0g sample for washing treatment, then place the bacteria in a 1.5mL centrifuge tube, immediately place it in liquid nitrogen to completely freeze, take it out and place it in a water bath at 65°C to slowly melt, repeat the freeze-thaw process 3 times, and then Add 0.1 mL of 10% SDS and 10.0 μl of 10 mg / mL proteinase K, shake in a constant temperature shaker at 37°C at 200 r / min for 2 h, centrifuge at 10,000 g for 10 min at room temperature, collect the supernatant and transfer it to another centrifuge tube. Add chloroform equal to the volume of the supernatant, centrifuge at 10,000 g for 10 min, absorb the supernatant and transfer it to another centrifuge tube for phenol-chloroform extraction twice, and finally transfer the supernatant to another centrifuge tube, add 2 times the supernatant Precipitate DNA with a clear v...

Embodiment 2

[0043] The method for identifying capsular rhodobacter comprises the steps:

[0044] 1. DNA extraction from photosynthetic bacteria products

[0045]Weigh 1.0g solid sample of photosynthetic bacteria (if it is a liquid sample, centrifuge the bacterial solution first, and take 1.0g of precipitate), then place the bacteria in a 1.5mL centrifuge tube, and immediately place it in liquid nitrogen to freeze completely. Place in a 65°C water bath to slowly thaw, and then freeze and thaw repeatedly 3 times, then add 0.1mL 10% SDS and 10.0μl 10mg / mL proteinase K, shake in a constant temperature shaker at 37°C at 200r / min for 2h, 10000g at room temperature After centrifugation for 10 min, the supernatant was collected and transferred to another centrifuge tube. Add the same volume of chloroform as the supernatant, mix well, centrifuge at 10000g for 10min, absorb the supernatant and transfer it to another centrifuge tube for phenol-chloroform extraction twice, finally transfer the super...

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Abstract

The invention relates to specific PCR primers for identifying rhodobacter capsulatus, a kit and application. The specific PCR primers comprise the following primer pairs: primer pairs shown in SEQ ID NO:1 and SEQ ID NO:2; primer pairs shown in SEQ ID NO:3 and SEQ ID NO:4; and primer pairs shown in SEQ ID NO:5 and SEQ ID NO:6. The specific PCR primers provided by the invention have high specificity to the rhodobacter capsulatus and solve the problem of poor capsule specificity in traditional methods. A method for identifying the rhodobacter capsulatus established by the primers has the advantages of good specificity, high stability and high sensitivity, can well control occurrence of false positive during detection and improves the detection accuracy of the rhodobacter capsulatus.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, to a specific PCR primer for identifying capsulated rhodobacter, a kit containing the primer and use. Background technique [0002] The pollution of the natural environment and the problem of antibiotic resistance in the process of animal breeding are becoming more and more serious. How to adopt new technologies and methods to promote health and ensure safe production has become the key to the sustainable development of human beings. It has also become a research hotspot at home and abroad. In fact, This is also a bottleneck for the further development of the bio-industry. In this context, the physiological and biochemical characteristics of Rhodobacter capsulata play an important role in human life and production, and have been widely studied and applied in water purification, aquaculture, crops, and horticulture. Chinese patent application number is 201610367225.X, a kind of pra...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11
CPCC12Q1/689C12Q1/686C12Q2565/125Y02A50/30
Inventor 周辉田云刘成国段希宇王晶晶
Owner HUNAN AGRICULTURAL UNIV
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